Attenuation of a Pathogenic Mycoplasma Strain by Modification of the obg Gene by Using Synthetic Biology Approaches
| Autor: | Pascal Sirand-Pugnet, Yanina Valverde Timana, Carole Lartigue, Brigitte Batailler, Sanjay Vashee, Joerg Jores, Fabien Labroussaa, Elise Schieck, Flavio Sacchini, Anne Liljander, Horst Posthaus, Alain Blanchard |
|---|---|
| Přispěvatelé: | Biologie du fruit et pathologie (BFP), Université Bordeaux Segalen - Bordeaux 2-Institut National de la Recherche Agronomique (INRA)-Université Sciences et Technologies - Bordeaux 1, Universität Bern- University of Bern [Bern], Partenaires INRAE, International Livestock Research Institute [CGIAR, Nairobi] (ILRI), International Livestock Research Institute [CGIAR, Ethiopie] (ILRI), Consultative Group on International Agricultural Research [CGIAR] (CGIAR)-Consultative Group on International Agricultural Research [CGIAR] (CGIAR), Institute of Veterinary Pathology, Justus-Liebig-Universität Gießen (JLU), J. Craig Venter Institute [La Jolla, USA] (JCVI), Lartigue, Carole |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
temperature sensitivity
Mutant lcsh:QR1-502 Virulence Mutagenesis (molecular biology technique) GTPase Obg Saccharomyces cerevisiae medicine.disease_cause Microbiology lcsh:Microbiology 03 medical and health sciences Mycoplasma medicine genome transplantation mollicute pathologie animale attenuated strain genome engineering vaccines caprin Molecular Biology Gene 030304 developmental biology 0303 health sciences [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health 630 Agriculture biology santé animale 030302 biochemistry & molecular biology biology.organism_classification Phenotype QR1-502 3. Good health Transplantation Médecine vétérinaire et santé animal 570 Life sciences Veterinary medicine and animal Health biologie synthétique bactérie pathogène Mycoplasma mycoides |
| Zdroj: | MSphere MSphere, American Society for Microbiology., 2019, 4 (3), pp.e00030-19. ⟨10.1128/mSphere.00030-19⟩ mSphere, Vol 4, Iss 3, p e00030-19 (2019) mSphere 3 (4), e00030-19. (2019) Lartigue, Carole; Valverde Timana, Yanina; Labroussaa, Fabien; Schieck, Elise; Liljander, Anne; Sacchini, Flavio; Posthaus, Horst; Batailler, Brigitte; Sirand-Pugnet, Pascal; Vashee, Sanjay; Jores, Jörg; Blanchard, Alain (2019). Attenuation of a Pathogenic Mycoplasma Strain by Modification of the obg Gene by Using Synthetic Biology Approaches. mSphere, 4(3) American Society for Microbiology 10.1128/mSphere.00030-19 mSphere, Vol 4, Iss 3 (2019) |
| ISSN: | 2379-5042 |
| DOI: | 10.1128/mSphere.00030-19⟩ |
| Popis: | Mycoplasma species are responsible for several economically significant livestock diseases for which there is a need for new and improved vaccines. Most of the existing mycoplasma vaccines are attenuated strains that have been empirically obtained by serial passages or by chemical mutagenesis. The recent development of synthetic biology approaches has opened the way for the engineering of live mycoplasma vaccines. Using these tools, the essential GTPase-encoding gene obg was modified directly on the Mycoplasma mycoides subsp. capri genome cloned in yeast, reproducing mutations suspected to induce a temperature-sensitive (TS+) phenotype. After transplantation of modified genomes into a recipient cell, the phenotype of the resulting M. mycoides subsp. capri mutants was characterized. Single-point obg mutations did not result in a strong TS+ phenotype in M. mycoides subsp. capri, but a clone presenting three obg mutations was shown to grow with difficulty at temperatures of ≥40°C. This particular mutant was then tested in a caprine septicemia model of M. mycoides subsp. capri infection. Five out of eight goats infected with the parental strain had to be euthanized, in contrast to one out of eight goats infected with the obg mutant, demonstrating an attenuation of virulence in the mutant. Moreover, the strain isolated from the euthanized animal in the group infected with the obg mutant was shown to carry a reversion in the obg gene associated with the loss of the TS+ phenotype. This study demonstrates the feasibility of building attenuated strains of mycoplasma that could contribute to the design of novel vaccines with improved safety. IMPORTANCE Animal diseases due to mycoplasmas are a major cause of morbidity and mortality associated with economic losses for farmers all over the world. Currently used mycoplasma vaccines exhibit several drawbacks, including low efficacy, short time of protection, adverse reactions, and difficulty in differentiating infected from vaccinated animals. Therefore, there is a need for improved vaccines to control animal mycoplasmoses. Here, we used genome engineering tools derived from synthetic biology approaches to produce targeted mutations in the essential GTPase-encoding obg gene of Mycoplasma mycoides subsp. capri. Some of the resulting mutants exhibited a marked temperature-sensitive phenotype. The virulence of one of the obg mutants was evaluated in a caprine septicemia model and found to be strongly reduced. Although the obg mutant reverted to a virulent phenotype in one infected animal, we believe that these results contribute to a strategy that should help in building new vaccines against animal mycoplasmoses. |
| Databáze: | OpenAIRE |
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